Fluid retention plates and analysis cartridges

ABSTRACT

Fluid storage containers and analysis cartridges for use in assay processes are presented. In addition, systems comprising such storage containers and analysis cartridges and methods of using such containers and cartridges are presented as well. In specific embodiments, fluid storage containers are configured to be coupled to analysis cartridges in a first stage and a second stage.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.15/427,432, filed Feb. 8, 2017, which is a continuation of U.S. patentapplication Ser. No. 14/169,284, filed Jan. 31, 2014, now U.S. Pat. No.9,597,685, which claims priority to U.S. Provisional Patent ApplicationSer. No. 61/759,210, filed Jan. 31, 2013, the contents of which areincorporated by reference herein.

FIELD OF THE INVENTION

This invention relates to bulk fluid storage containers configured foruse with printed circuit boards (PCB) or “labs on a chip” to performfluid assays. In particular, this invention relates to bulk fluidstorage containers that are configured to be disposable (in other words,are configured for a single use) and are further configured to bepartially or fully pre-loaded with reagents, oils, or other fluids forperforming a fluid assay.

BACKGROUND

The following descriptions and examples are not admitted to be prior artby virtue of their inclusion within this section.

Fluid assays are used for a variety of purposes, including but notlimited to biological screenings and environmental assessments.Sometimes, fluid assays may need to be performed in the field away froma laboratory. In the field environment it is useful to have a systemthat is capable of performing the entire assay from sample preparationto analysis (sample to answer) without the use of conventionallaboratory tools. This type of system may, for example, take in a userinput sample and process the sample using reagents, heat, magneticparticles, and/or imaging to produce an assay result for the sample.

SUMMARY OF THE INVENTION

Fluid storage containers and analysis cartridges for use in assayprocesses are presented. In addition, systems comprising such storagecontainers and analysis cartridges and methods of using such containersand cartridges are presented. In specific embodiments, fluid storagecontainers are configured to be coupled to analysis cartridges in afirst stage and a second stage.

Exemplary embodiments include a system comprising a first plate and asecond plate coupleable to the first plate. In certain embodiments, thefirst plate may comprise: a first side; a second side; a first reservoircoupled to the first side of the first plate; a protrusion extendingfrom the second side of the first plate; a first channel extending fromthe first side of the first plate through the protrusion on the secondside of the first plate; a fluid displacement mechanism in fluidcommunication with the first reservoir and the first channel; and apierceable seal coupled to the protrusion extending from the second sideof the first plate and covering the first channel. In specificembodiments, the second plate may comprise: a first side; a second side;a protrusion extending from the first side of the second plate andconfigured to mate with the protrusion on the second side of the firstplate; a piercing element associated with the protrusion extending fromthe first side of the second plate, where the piercing element isconfigured to extend through the pierceable seal when the protrusion onthe second side of the first plate is mated with the protrusionextending from the first side of the second plate; and a second channelin fluid communication with the protrusion extending from the first sideof the second plate.

In particular embodiments, the fluid displacement mechanism may be apiston. In certain embodiments, the fluid displacement mechanism may bea flexible blister. In specific embodiments, the first plate may furthercomprise a sample fluid displacement mechanism and a sample inputchannel configured to conduct a fluid from the first side of the firstplate to the sample fluid displacement mechanism. Particular embodimentsmay also comprise a removable sample input cap coupled to a samplereservoir. Certain embodiments may further comprise a gasket coupled tothe second side of the first plate, where the gasket is configured toform a substantially fluid-tight seal between the protrusion extendingfrom the first side of the second plate and the protrusion extendingfrom the second side of the first plate.

Specific embodiments may further comprise an assay surface coupled tothe second side of the second plate. In particular embodiments, theassay surface may comprise an electrowetting surface. In certainembodiments, the assay surface may comprise microfabricated channelsand/or one or more heating elements.

In specific embodiments, the second plate may comprise a vent extendingfrom the first side of the second plate to the second side of the secondplate. Certain embodiments may further comprise a hydrophobic nylon meshcoupled to at least a portion of the first side of the second plate andcovering the vent. In particular embodiments, the second side of thefirst plate may be coupled to the first side of the second plate.

In specific embodiments, the first plate may comprise: a plurality ofprotrusions extending from the second side of the first plate; aplurality of reservoirs; a plurality of channels extending from thefirst side of the first plate through the plurality of protrusions onthe second side of the first plate; a plurality of fluid displacementmechanisms, where each fluid displacement mechanism is in fluidcommunication with a reservoir and a channel extending from the firstside of the first plate through the plurality of protrusions on thesecond side of the first plate. In certain embodiments, the first platemay also comprise a plurality of pierceable seals, where each pierceableseal in the plurality of pierceable seals is coupled to one of theprotrusions in the plurality of protrusions extending from the secondside of the first plate, and where each pierceable seal covers at leasta portion of one of the channels in the plurality of channels.

In particular embodiments, the second plate may comprise: a plurality ofprotrusions extending from the first side of the second plate, eachprotrusion of which is respectively configured to mate with one of theprotrusions in the plurality of protrusions on the second side of thefirst plate. In specific embodiments, the second plate may comprise aplurality of piercing elements, where each piercing element isconfigured to pierce one of the pierceable seals in the plurality ofpierceable seals when the protrusions on the second side of the firstplate are respectively mated with the protrusions extending from thefirst side of the second plate. In certain embodiments, the second platemay comprise a plurality of channels extending from the second side ofthe second plate through the plurality of piercing elements disposedwithin the protrusions extending from the first side of the secondplate.

In particular embodiments, the first plate may have a thickness between0.05 inches and 0.5 inches. In specific embodiments, the second platemay have a thickness between 0.05 inches and 0.5 inches. In certainembodiments, the first plate may be a rigid plastic plate. In particularembodiments the second plate may be a rigid plastic plate.

Embodiments of fluid storage containers are disclosed. In certainembodiments, a fluid storage container may be configured to be coupledto an analysis cartridge. The fluid storage container may comprise: abulk fluids plate comprising: at least one reagent reservoir unitcomprising: a barrel comprising a channel; a pierceable seal coupled tothe barrel; and a fluid displacement mechanism in fluid communicationwith the channel; where each reagent reservoir unit is configured tocontain a volume of fluid. In various embodiments, each barrel may beconsidered a protrusion extending from a side (e.g., the bottom side) ofthe bulk fluids plate.

In certain embodiments, the bulk fluids plate has a bottom side, and thebulk fluids plate may further comprise a first pair of tabs and a secondpair of tabs, where the first pair of tabs is longer than the secondpair of tabs, and each tab extends from the bottom side. In particularembodiments, the bulk fluids plate also includes a sample reservoirunit.

Particular embodiments may comprise a sample dispensing channel in fluidcommunication with the sample reservoir. In specific embodiments, thesample reservoir may comprise: a removable sample input cap; a sampleinput channel; and a sample fluid displacement mechanism in fluidcommunication with the sample input channel. In particular embodiments,the bulk fluids plate has a top side, and the removable sample input capmay be located on the top side, and the sample fluid displacementmechanism may be located on the bottom side. In certain embodiments, thebulk fluids plate may further comprise a sample control reservoirconfigured to retain a sample control. In particular embodiments, thebulk fluids plate may further comprise a lysis fluid displacementmechanism in fluid communication with a volume of lysate, a lysis fluiddisplacement mechanism in fluid communication with a volume of lysate.In particular embodiments, the lysate is configured to be delivered tothe sample control reservoir when the lysis fluid displacement mechanismis actuated.

In certain embodiments, the sample reservoir unit may be in fluidcommunication with the sample control reservoir via a lysis channel. Inparticular embodiments, the bulk fluids plate may comprise a gasketcoupled to at least one protrusion. In specific embodiments, the bulkfluids plate may comprise a plurality of protrusions, and the gasket maycomprise a plurality of holes equal or greater in number to the numberof protrusions in the plurality of protrusions. In certain embodiments,each reagent reservoir unit may contain a volume of fluid selected fromthe group consisting of oil, imaging dilution buffer, binding beads,binding buffer, wash buffer, rehydration buffer, and lysis buffer.

In particular embodiments, each reagent reservoir unit may comprisebetween about 20 uL and about 20 mL of fluid. In specific embodiments,each reagent reservoir unit may comprise between about 20 uL and about20 mL of fluid. In certain embodiments, the bulk fluids plate maycomprise eight reagent reservoir units. In particular embodiments, aprotrusion may comprise a distal end distal from the fluid displacementmechanism and the pierceable seal may be located on the distal end ofthe protrusion. In specific embodiments, the pierceable seal maycomprise foil. In certain embodiments, the bulk fluids plate maycomprise plastic.

Certain embodiments include a fluid storage container comprising: asyringe barrel array comprising: multiple isolated reservoirs in asingle part or multiple individual reservoirs in separate partscomprising: a barrel comprising a channel; a pierceable seal coupled tothe barrel; and a piston or some other means to achieve a fluid tightseal that when actuated results in a positive displacement pump action,covering a portion of the channel; where each reagent reservoir unit isconfigured to contain a volume of fluid.

In specific embodiments, a fluid storage container configured to becoupled to an analysis cartridge is disclosed, the fluid storagecontainer comprising: a bulk fluids plate comprising: at least onereagent reservoir unit comprising: a barrel comprising a channel; apierceable seal coupled to the barrel; and a fluid displacementmechanism (including e.g. a piston or flexible blister) covering atleast a portion of the channel; where each reagent reservoir unit isconfigured to contain a volume of fluid. In various embodiments, eachbarrel may be considered a protrusion extending from a side (e.g., thebottom side) of the bulk fluids plate.

Another embodiment may be a fluid storage container comprising: asyringe barrel array comprising: multiple isolated reservoirs in asingle part or multiple individual reservoirs in separate partscomprising: a barrel comprising a channel; a pierceable seal coupled tothe barrel; and a piston or some other means to achieve a fluid tightseal that when actuated results in a positive displacement pump action,covering a portion of the channel; where each reagent reservoir unit isconfigured to contain a volume of fluid.

In other embodiments, an analysis cartridge may be configured to becoupled to a fluid storage container. In certain embodiments, theanalysis cartridge may comprise: a retention plate comprising: a topside and a bottom side; a plurality of bosses, each boss comprising: achamber having a floor; and a lance extending from the floor andcomprising a conduit through the floor; where each boss is configured toreceive a barrel and each lance is configured to pierce a pierceableseal on the barrel; and a plurality of dispensing reservoirs, eachdispensing reservoir in fluid communication with a conduit of one of theplurality of bosses. In various embodiments, each boss may be considereda protrusion extending from a side (e.g., the top side) of the retentionplate.

In certain embodiments, the retention plate may comprise a plurality ofslots extending between the top side and the bottom side. Particularembodiments may comprise a printed circuit board (PCB) coupled to thebottom side of the retention plate. In specific embodiments, each lancemay be between about 0.005″ tall and about 0.080″ tall. In certainembodiments, bosses in the plurality of protrusions may be disposed onthe top side of the retention plate.

In still other embodiments, a system is disclosed comprising: a firstplate comprising: a first side; a second side; a protrusion extendingfrom the second side of the first plate; a first channel extending fromthe first side of the first plate through the protrusion on the secondside of the first plate; a fluid displacement mechanism (e.g. a pistonor a flexible blister) coupled to the first side of the first plate anddefining a volume over the first channel; and a pierceable seal coupledto the protrusion extending from the second side of the first plate andcovering the first channel; and a second plate coupleable to the firstplate comprising: a first side; a second side; a protrusion extendingfrom the first side of the second plate and configured to receive theprotrusion on the second side of the first plate; a piercing elementdisposed within the protrusion extending from the first side of thesecond plate, where the piercing element is configured to pierce thepierceable seal when the protrusion on the second side of the firstplate is disposed in the protrusion extending from the first side of thesecond plate; and a second channel extending from the second side of thesecond plate through the piercing element disposed within the protrusionextending from the first side of the second plate.

In certain embodiments, a flexible blister may be coupled to the firstplate with an adhesive. Particular embodiments may comprise a sampleblister coupled to the second side of the first plate and a sample inputchannel configured to conduct a fluid from the first side of the firstplate to the sample blister. Specific embodiments may comprise aremovable sample input cap coupled to the first side of the first plateand covering the sample input channel. Certain embodiments may comprisea gasket coupled to the second side of the first plate, where the gasketis configured to form a substantially fluid-tight seal between theprotrusion extending from the first side of the second plate and thesecond side of the first plate. Particular embodiments may comprise agasket coupled to the second side of the first plate, where the gasketis configured to form a substantially fluid-tight seal between theprotrusion extending from the first side of the second plate and thesecond side of the first plate. Specific embodiments may comprise agasket coupled to the second side of the first plate, where the gasketis configured to form a substantially fluid-tight seal between theprotrusion extending from the first side of the second plate and thesecond side of the first plate. In particular embodiments, the assaysurface may comprise an electrowetting surface, microfabricatedchannels, and/or microfabricated channels.

In certain embodiments, the second plate may comprise a vent extendingfrom the first side of the second plate to the second side of the secondplate. Particular embodiments may comprise a hydrophobic nylon meshcoupled to at least a portion of the first side of the second plate andcovering the vent. In specific embodiments the second side of the firstplate may be coupled to the first side of the second plate. In certainembodiments the first plate may comprise: a plurality of protrusionsextending from the second side of the first plate; a plurality ofchannels extending from the first side of the first plate through theplurality of protrusions on the second side of the first plate; aplurality of flexible blisters coupled to the first side of the firstplate, where each blister of the plurality of blisters at leastpartially covers at least one channel of the plurality of channels; anda plurality of pierceable seals, where each pierceable seal in theplurality of pierceable seals is coupled to one of the protrusions inthe plurality of protrusions extending from the second side of the firstplate, and where each pierceable seal covers at least a portion of oneof the channels in the plurality of channels.

In particular embodiments, the second plate may comprise: a plurality ofprotrusions extending from the first side of the second plate, eachprotrusion of which is respectively configured to mate with one of theprotrusions in the plurality of protrusions on the second side of thefirst plate; a plurality of piercing elements, where each piercingelement is configured to pierce one of the pierceable seals in theplurality of pierceable seals when the protrusions on the second side ofthe first plate are respectively mated with the protrusions extendingfrom the first side of the second plate; and a plurality of channelsextending from the second side of the second plate through the pluralityof piercing elements disposed within the protrusions extending from thefirst side of the second plate.

In specific embodiments, the first plate may have a thickness between0.05 inches and 0.5 inches. In some embodiments, the second plate mayhave a thickness between 0.05 inches and 0.5 inches. In certainembodiments, the first plate and/or the second plate may be a rigidplastic plate.

In other embodiments, a system is disclosed comprising: a fluid storagecontainer comprising: a bulk fluids plate comprising: a top side and abottom side; a sample reservoir configured to receive a sample volume; aplurality of reagent reservoirs, each of which comprises a fluid volume;and a first pair of tabs and a second pair of tabs, where each tab inthe first pair of tabs is longer than each tab in the second pair oftabs, and the tabs extend from the bottom side; and an analysiscartridge coupleable to the fluid storage container, the analysiscartridge comprising: a retention plate comprising: a top side and abottom side; a plurality of bosses, each boss comprising a chamber, afloor, and a conduit through the floor: a plurality of dispensingreservoirs, each dispensing reservoir in fluid communication with aconduit of one of the bosses; and a plurality of slots, each slot beingsized and positioned to respectively receive one of the tabs in thefirst and second pairs of tabs when the analysis cartridge and the fluidstorage container are coupled to each other in an engaged state, thesystem being configured such that in an intermediate stage of coupling,fewer than all of the tabs extend respectively through all of the slots.

In particular embodiments, the analysis cartridge may comprise ananalysis element coupled to the retention plate. In some embodiments,the fluid storage container may be configured to be coupled to theanalysis cartridge in a first stage and a second stage. In specificembodiments, the fluid storage container may be coupled to the analysiscartridge with the plurality of pierceable seals intact in the firststage; and the fluid storage container may be coupled to the analysiscartridge with at least one pierceable seal pierced by the analysiscartridge in the second stage. In certain embodiments, each flexibleblister of each reagent reservoir unit may project from the top side ofthe bulk fluids plate. In particular embodiments, the fluid storagecontainer may further comprise a first pair of tabs and a second pair oftabs, where the first pair of tabs is longer than the second pair oftabs and the tabs extend from the bottom side of the fluid storagecontainer. In specific embodiments, the analysis cartridge may furthercomprise a plurality of slots, and at least four of the slots may beconfigured to receive a tab of the fluid storage container.

In particular embodiments, the fluid storage container may comprise asample reservoir unit. In certain embodiments, the sample reservoir unitmay comprise a removable sample input cap, a removable sample input cap,and a sample blister. In some embodiments, the fluid storage containermay be coupled to the analysis cartridge in the first stage. In specificembodiments, each reagent reservoir unit may contain a volume of fluidselected from the group consisting of oil, imaging dilution buffer,binding beads, binding buffer, wash buffer, rehydration buffer, andlysis buffer. In certain embodiments, the fluid storage container mayfurther comprise a frangible lysis blister configured to contain avolume of lysis buffer. In particular embodiments, the fluid storagecontainer may comprise a sample control. In some embodiments, theanalysis cartridge may comprise a printed circuit board coupled to theretention plate.

In still other embodiments a method of preparing an assay is disclosedcomprising obtaining a system comprising a fluid storage container andan analysis cartridge coupleable to the fluid storage container. In someembodiments, the fluid storage container may comprise: a bulk fluidsplate comprising: a top side and a bottom side; a sample reservoirconfigured to receive a sample volume; a plurality of reagentreservoirs, each of which comprises a fluid volume; and a first pair oftabs and a second pair of tabs, where each tab in the first pair of tabsis longer than each tab in the second pair of tabs, and the tabs extendfrom the bottom side. In some embodiments, the analysis cartridge maycomprise: a retention plate comprising: a top side and a bottom side; aplurality of bosses, each boss comprising a chamber, a floor, and aconduit through the floor: a plurality of dispensing reservoirs, eachdispensing reservoir in fluid communication with a conduit of one of thebosses; and a plurality of slots, each slot being sized and positionedto respectively receive one of the tabs in the first and second pairs oftabs when the analysis cartridge and the fluid storage container arecoupled to each other in an engaged state, the system being configuredsuch that in an intermediate stage of coupling, fewer than all of thetabs extend respectively through all of the slots. In particularembodiments, the method may also comprise adding a volume of sample tothe sample reservoir; and coupling the fluid storage container to theanalysis cartridge in the first stage.

In certain embodiments, the fluid storage container may be coupled tothe analysis cartridge prior to adding the volume of sample to thesample reservoir unit. In particular embodiments, the fluid storagecontainer may be coupled to the analysis cartridge in a first stage withthe pierceable seals intact prior to adding the volume of sample to thesample reservoir unit. Certain embodiments may comprise coupling thefluid storage container to the analysis cartridge in a second stage withat least one of the pierceable seals pierced. In specific embodiments,the fluid storage container may be coupled to the analysis cartridgeafter adding the volume of sample to the sample reservoir unit.

Particular embodiments may comprise actuating the flexible blister of atleast one of the plurality of reagent reservoir units to deliver thevolume of fluid in the reagent reservoir unit to the analysis elementvia the conduit in the lance. Specific embodiments may compriseactuating two or more of the flexible blisters simultaneously. Certainembodiments may comprise actuating two or more of the flexible blisterssequentially. In particular embodiments, at least one of the pluralityof reagent reservoir units comprises a volume of oil, and the flexibleblister of the at least one reagent reservoir unit comprising a volumeof oil may be actuated before actuating the flexible blister of any ofthe other reagent reservoir units and before actuating the sampleblister.

In certain embodiments, the fluid storage container further comprises afrangible lysis blister comprising a lysis buffer, and the method maycomprise actuating frangible lysis blister such that lysis buffer isdistributed to the volume of sample.

In various embodiments of the disclosed apparatuses, systems, andmethods, the fluid storage container may comprise one, two, three, four,five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen,fifteen, sixteen, seventeen, eighteen, nineteen, twenty, or more reagentreservoir units. Also in such disclosed embodiments, the analysiscartridge may comprise one, two, three, four, five, six, seven, eight,nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen,seventeen, eighteen, nineteen, twenty, or more bosses (or protrusions).In preferred embodiments, the number of bosses (or protrusions) is equalto the total number of reagent reservoir units and sample reservoirunits.

In certain embodiments of the disclosed apparatuses, systems, andmethods, the fluid storage container may come preloaded with fluids inone or more of the reagent reservoir units for performing a fluid assay.The reagent reservoirs may comprise oil, imaging dilution buffer,binding beads, binding buffer, wash buffer, rehydration buffer, andlysis buffer in various embodiments. In specific embodiments, eachreagent reservoir unit may comprise between about 20 uL and about 20 mL,between about 20 uL and about 2 mL, between about 20 uL and about 1 mL,between about 20 uL and about 200 uL, between about 50 uL and about 2mL, or between about 10 uL and about 200 uL of fluid.

In various embodiments of the disclosed apparatuses, systems, andmethods, the fluid storage container comprises a first tab and a secondtab, where the first tab and second tab are of different lengths. Inpreferred embodiments, the tabs are paired such that there are at leasttwo tabs of a substantially the same length. In still other embodiments,the longer tabs may be considered fastening tabs and the shorter tabsmay be considered engaging tabs. In such embodiments, there may be one,two, three, four, five, or more pairs of fastening tabs and one, two,three, four, five, or more pairs of engaging tabs.

In addition, in various embodiments of the disclosed apparatuses,systems, and methods, the analysis cartridge may comprise slots throughthe top and bottom of the retention plate configured to receive andequal or greater in number to the total number of tabs. So, in variousembodiments, the analysis cartridge may comprise one, two, three, four,five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen,fifteen, sixteen, seventeen, eighteen, nineteen, twenty, or more slots.

In alternate embodiments of the disclosed apparatuses, systems andmethods, the slots may be located on the fluid storage container and thetabs may be located on the analysis cartridge.

In alternate embodiments of the disclosed apparatuses additionalfasteners such as threaded features can be used to engage the fluidstorage container to the analysis cartridge.

In certain embodiments of the disclosed apparatuses, systems, andmethods, each boss of the analysis cartridge comprises a lance, theheight of which may be about 0.100″, 0.120″, 0.140″, 0.160″, 0.180″,0.200″, 0.220″, 0.240″, or 0.260″.

In some embodiments of the disclosed apparatuses, systems, and methods,the fluid storage container and/or the analysis cartridge furthercomprises a gasket configured to form a substantially leak-proof sealwith at least a portion of the analysis cartridge when the fluid storagecontainer is coupled to the analysis cartridge in the second stage. Incertain specific embodiments, the gasket may comprise a plurality ofholes equal or greater in number to the total number of the plurality ofbarrels or bosses, that is, the gasket may comprise one, two, three,four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen,fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, ormore holes.

Embodiments of the disclosed fluid storage containers and analysiscartridges may comprise polycarbonate, polyurethane, polyester, epoxyresin, phenolic resin, polyethylene (PE), polypropylene (PP), polyvinylchloride (PVC), or polyethylene terephthalate (PET or PETE).

The term “coupled” is defined as connected, although not necessarilydirectly, and not necessarily mechanically. Two items are “coupleable”if they can be coupled to each other, and, when coupled, may still becharacterized as “coupleable.” Unless the context explicitly requiresotherwise, items that are coupleable are also decoupleable, andvice-versa. One non-limiting way in which a first structure iscoupleable to a second structure is for the first structure to beconfigured to be coupled (or configured to be coupleable) to the secondstructure.

The terms “a” and “an” are defined as one or more unless this disclosureexplicitly requires otherwise.

The term “substantially” and its variations (e.g., “approximately” and“about”) are defined as being largely but not necessarily wholly what isspecified (and include wholly what is specified) as understood by one ofordinary skill in the art. In any disclosed embodiment, the terms“substantially,” “approximately,” and “about” may be substituted with“within [a percentage] of” what is specified, where the percentageincludes 0.1, 1, 5, and 10 percent.

The terms “comprise” (and any form of comprise, such as “comprises” and“comprising”), “have” (and any form of have, such as “has” and“having”), “include” (and any form of include, such as “includes” and“including”) and “contain” (and any form of contain, such as “contains”and “containing”) are open-ended linking verbs. As a result, a method ordevice that “comprises,” “has,” “includes” or “contains” one or moresteps or elements possesses those one or more steps or elements, but isnot limited to possessing only those one or more elements. Likewise, astep of a method or an element of a device that “comprises,” “has,”“includes” or “contains” one or more features possesses those one ormore features, but is not limited to possessing only those one or morefeatures. For example, a fluid retention plate that comprises a samplereservoir unit has one sample reservoir unit, but may have more than onesample reservoir unit.

Furthermore, a device or structure that is configured in a certain wayis configured in at least that way, but may also be configured in waysthat are not listed. Metric units may be derived from the English unitsprovided by applying a conversion and rounding to the nearestmillimeter.

The feature or features of one embodiment may be applied to otherembodiments, even though not described or illustrated, unless expresslyprohibited by this disclosure or the nature of the embodiments.

Any embodiment of any of the disclosed devices and methods can consistof or consist essentially of—rather thancomprise/include/contain/have—any of the described elements and/orfeatures and/or steps. Thus, in any of the claims, the term “consistingof” or “consisting essentially of” can be substituted for any of theopen-ended linking verbs recited above, in order to change the scope ofa given claim from what it would otherwise be using the open-endedlinking verb.

Other features and associated advantages will become apparent withreference to the following detailed description of specific embodimentsin connection with the accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

The following drawings illustrate by way of example and not limitation.For the sake of brevity and clarity, every feature of a given structuremay not be labeled in every figure in which that structure appears.Identical reference numbers do not necessarily indicate an identicalstructure. Rather, the same reference number may be used to indicate asimilar feature or a feature with similar functionality, as maynon-identical reference numbers.

The embodiments of the present fluid storage containers and analysiscartridges shown in FIGS. 1-10B are drawn to scale.

FIG. 1 is a top perspective view of embodiments of an assembly of afluid storage container and an analysis cartridge.

FIG. 2 is a bottom perspective view of embodiments of an assembly of afluid storage container and an analysis cartridge.

FIG. 3 is an exploded view of embodiments of a fluid storage containerand an analysis cartridge.

FIG. 4 is a top perspective view of an embodiment of a bulk fluidsplate.

FIG. 5 is a bottom perspective view of an embodiment of a bulk fluidsplate with pierceable seals removed.

FIG. 6 is a bottom perspective view of an embodiment of a bulk fluidsplate with pierceable seals shown.

FIG. 7 is a top perspective view of an embodiment of a gasket.

FIG. 8 is a top view of an embodiment of a retention plate.

FIG. 9 is a perspective view of embodiments of a PCB.

FIG. 10A is a side section view of embodiments of fluid storagecontainer coupled to analysis cartridge at the fastened stage.

FIG. 10B is a side section view of embodiments of fluid storagecontainer coupled to analysis cartridge at the engaged stage

FIG. 11 is a top perspective view of embodiments of an assembly of afluid storage container and an analysis cartridge.

FIG. 12 is a bottom perspective view of embodiments of an assembly of afluid storage container and an analysis cartridge.

FIG. 13 is an exploded view of embodiments of a fluid storage containerand an analysis cartridge.

FIG. 14 is a side section view of embodiments of a fluid storagecontainer and an analysis cartridge.

FIG. 15 is a top perspective view of an embodiment of a bulk fluidsplate.

FIG. 16 is a bottom perspective view of an embodiment of a bulk fluidsplate.

FIG. 17 is a bottom perspective view of an embodiment of a bulk fluidsplate with pierceable seals shown.

FIG. 18 is a bottom perspective view of an embodiment of a fluiddisplacement mechanism.

FIG. 19 is a top perspective view of an embodiment of a gasket.

DETAILED DESCRIPTION

Various features and advantageous details are explained more fully withreference to the non-limiting embodiments that are illustrated in theaccompanying drawings and detailed in the following description. Itshould be understood, however, that the detailed description and thespecific examples, while indicating embodiments of the invention, aregiven by way of illustration only, and not by way of limitation. Varioussubstitutions, modifications, additions, and/or rearrangements willbecome apparent to those of ordinary skill in the art from thisdisclosure.

In the following description, numerous specific details are provided toprovide a thorough understanding of the disclosed embodiments. One ofordinary skill in the relevant art will recognize, however, that theinvention may be practiced without one or more of the specific details,or with other methods, components, materials, and so forth. In otherinstances, well-known structures, materials, or operations are not shownor described in detail to avoid obscuring aspects of the invention.

FIGS. 1 and 2 are top and bottom perspective views, respectively, ofsystem 5—one embodiment of the present systems—which comprises fluidstorage container 10 (one embodiment of the present containers) coupledto analysis cartridge 20 (one embodiment of the present cartridges).FIG. 3 is an exploded view of fluid storage container 10 and analysiscartridge 20.

While container 10 and cartridge 20 may be sold or provided in system 5,they may also be sold or provided separately, as may other embodimentsof the present systems, containers, and cartridges.

As discussed in more detail below, fluid storage container 10 comprisesa plurality of reservoir units that may be preloaded with liquidreagents, oils, or other fluids usable to perform an assay. Fluidstorage container 10 may comprise reservoir units that contain assaycomponents that are not in fluidic form, such as lyophilized, dried, orpowdered reagents. In the embodiments shown, the fluid assay isperformed by an analysis element, such as the printed circuit board(PCB) 800, which is coupled to analysis cartridge 20. In otherembodiments, other suitable analysis elements may be used.

For example, certain embodiments of the present fluid storage containersare configured to be coupled to an analysis cartridge in at least twostages. Turning to system 5 specifically, in the first or “fastened”stage, fluid storage container 10 is secured to analysis cartridge 20such that fluid storage container 10 is not readily separable fromanalysis cartridge 20. In the second or “engaged” stage, fluid storagecontainer 10 is secured to analysis cartridge 20 such that lances oncartridge 20 pierce seals on the reservoirs containing the fluids. Afluid-tight (or substantially fluid-tight) path is thereby createdbetween each reservoir and the PCB. Each reservoir unit may besequentially engaged such that fluid is forced from the reservoir, alongthe fluid path, and onto the PCB to perform a fluid assay.

Fluid Storage Container

As shown in FIGS. 1-7, and especially in FIG. 3, fluid storage container10 comprises a bulk fluids plate 100 coupled to a blister layer 130 viaan adhesive layer 140. Bulk fluids plate 100 comprises a plurality ofbarrels 111-119 (see FIG. 5) protruding downward from the underside ofbulk fluids plate 100. These barrels may also be characterized asprotrusions. In the embodiment shown in FIG. 7, gasket 200 is configuredto be coupled to bulk fluids plate 100 such that the plurality of holes201-209 in gasket 200 mate with the plurality of barrels 111-119. In theillustrated embodiment, barrels 111-119 are substantially cylindrical inshape (in other words, they have a substantially circular crosssection), but in other embodiments the barrels have other shapes (andshapes that differ from each other) such as substantially triangular,square, pentagonal, hexagonal, heptagonal, octagonal, or other polygonalcross sections, may be elliptical or partially rounded, or may haveirregular or fanciful cross sections.

As shown in FIGS. 1-4, bulk fluids plate 100 comprises a samplereservoir unit 150 that is configured to receive, retain, and dispense avolume of a sample. Sample reservoir unit 150 is bounded by a removablesample input cap 151, a sample input channel 152, and a sample blister153. In the illustrated embodiment, sample input port 155 is disposed ontop of bulk fluids plate 100 and comprises a sample input channel 152,which through bulk fluids plate 100. Sample input cap 151 is coupled toa sample input port 155, such as with a Luer lock connection. Sampleblister 153 is coupled to the underside of bulk fluids plate via anadhesive layer 141.

Bulk fluids plate 100 also comprises a sample control reservoir 160 (seeFIG. 4) within a sample control barrel 162, the sample control reservoir160 being configured to retain a sample control (such as a lyosphere,not pictured). Frangible lysis blister 161 is located adjacent to samplecontrol reservoir 160 such that tab 163 of lysis blister 161 is abovesample control reservoir 160 and between locating features 188. Tab 163seals the tops of the sample control reservoir 160 such that the samplecontrol lyosphere is retained within it. In certain embodiments,frangible lysis blister 161 comprises about 0.5 mL of lysis buffer. Inthe embodiment shown, when frangible lysis blister 161 is actuated,fluid within the blister is configured to rehydrate the sample controlin sample control reservoir 160.

Sample control reservoir 160 is in fluid communication with samplereservoir 150 through lysis channel 165 and lysis port 167. Lysischannel 165 is sealed by blister layer 130.

As shown in FIGS. 1-6 bulk fluids plate 100 further comprises a sampledispensing gutter 193 and a sample dispensing barrel 119 comprising asample dispensing channel 192. Sample dispensing gutter 193 is sealed byblister layer 130 and is in fluid communication with sample reservoir150 via proximal port 194. Sample dispensing gutter 193 is in fluidcommunication with sample dispensing channel 192. Sample dispensingchannel 192 is sealed by a pierceable sample seal 309 coupled to barrel119. In the embodiment shown in FIGS. 1, 2, and 4-6, fluid is configuredto travel from sample reservoir 150 through proximal port 194, throughsample dispensing gutter 193, and to sample dispensing channel 192. Whenpierceable sample seal 309 is pierced, fluid is allowed to selectivelyflow to the PCB.

Fluid storage container 10 further comprises a plurality of reagentreservoirs. The illustrated embodiment comprises a first reagentreservoir unit 101, a second reagent reservoir unit 102, a third reagentreservoir unit 103, a fourth reagent reservoir unit 104, a fifth reagentreservoir unit 105, a sixth reagent reservoir unit 106, a seventhreagent reservoir unit 107, and an eighth reagent reservoir unit 108.Fluid storage container 10 may comprise a greater or fewer number ofreagent reservoirs units in other embodiments depending on the assayparameters. Note that the term “reagent reservoir unit” and relatedterms are not strictly limited to reservoir units that contain reagents,which are substances for use in a chemical reaction; other fluids usedin assay preparation may be contained within reagent reservoirs, such asoils that are used to displace air from cartridge 20.

In the illustrated embodiment, each reagent reservoir is a sealed spacedefined by a barrel comprising a reagent channel, the channel beingsealed at the bottom end by a pierceable seal and sealably covered atthe top by a flexible blister. Barrels are located on the bottom side ofbulk fluids plate 100 (see FIGS. 5 and 6), while flexible blisters andblister layer 130 are located at the top side of bulk fluids plate 100(shown in FIG. 4). Accordingly, as shown in FIGS. 5 and 6 and in thecross section detail of FIGS. 10A and 10B, first reagent reservoir unit101 comprises first barrel 111 comprising a first reagent channel 121, afirst pierceable seal 301 coupled to the first barrel, and a firstflexible blister 131 in fluid communication with first channel 121. Inthe illustrated embodiment, each flexible seal is located at the distalend of each barrel. In other embodiments, each flexible seal may belocated within each barrel.

The other reagent reservoirs comprise similar features and areconfigured in a similar manner. Second reagent reservoir unit 102comprises second barrel 112 comprising a second reagent channel 122, asecond pierceable seal 302 coupled to second barrel 112, and a secondflexible blister 132 covering second reagent channel 122. Third reagentreservoir unit 103 comprises third barrel 113 comprising a third reagentchannel 123, a third pierceable seal 303 coupled to third barrel 113,and a third flexible blister 133 covering third reagent channel 123.Fourth reagent reservoir unit 104 comprises fourth barrel 114 comprisinga fourth reagent channel 124, a fourth pierceable seal 304 coupled tofourth barrel 114, and a fourth flexible blister 134 covering fourthreagent channel 124. Fifth reagent reservoir unit 105 comprises fifthbarrel 115 comprising a fifth reagent channel 125, a fifth pierceableseal 305 coupled to fifth barrel 115, and a fifth flexible blister 135covering fifth reagent channel 125. Sixth reagent reservoir unit 106comprises sixth barrel 116 comprising a sixth reagent channel 126, asixth pierceable seal 306 coupled to sixth barrel 116, and a sixthflexible blister 136 covering sixth reagent channel 126. Seventh reagentreservoir unit 107 comprises seventh barrel 117 comprising a seventhreagent channel 127, a seventh pierceable seal 307 coupled to seventhbarrel 117, and a seventh flexible blister 137 covering seventh reagentchannel 127. Eighth reagent reservoir unit 108 comprises eighth barrel118 comprising an eighth reagent channel 128, an eighth pierceable seal308 coupled to an eighth barrel 118, and an eighth flexible blister 138covering eighth reagent channel 128.

In a specific embodiment, first reagent reservoir unit 101 comprisesabout 70 uL of an imaging dilution buffer; second reagent reservoir unit102 comprises about 70 uL of a wash buffer; third reagent reservoir unit103 comprises about 70 uL of binding beads; fourth reagent reservoirunit 104 comprises about 0.6 mL of oil; fifth reagent reservoir unit 105comprises about 0.5 mL of oil; sixth reagent reservoir unit 106comprises about 280 uL of binding buffer; seventh reagent reservoir unit107 comprises about 70 uL of wash buffer; and eighth reagent reservoirunit 108 comprises about 70 uL of rehydration buffer. In otherembodiments, reagent reservoir units may comprise other fluids useful inperforming a fluid assay. In addition, other embodiments may comprisereagent reservoirs capable of containing a larger or smaller volume offluid. In specific embodiments, no reagent reservoir unit is configuredto contain about (and in more specific embodiments, no more than) 2 mLof fluid. In other specific embodiments, each reagent reservoir unit isconfigured to contain more than about (and, in more specificembodiments, more than) 20 uL of fluid.

As shown in FIGS. 1-6, bulk fluids plate 100 further comprise pairs oftabs located on opposite sides of the plate. In the embodiment shown,there are two fastening tabs 182 and four engagement tabs 184. Fasteningtabs 182 are longer than the engagement tabs 184 to facilitate securingfluid storage container 10 to analysis cartridge 20 in two stages. Thefastening tabs 182 and the engagement tabs 184 are paired in theillustrated embodiment, but may be not paired in other embodiments.

As shown in FIGS. 1, 2, 3, and 7, the illustrated embodiment of fluidstorage container 10 further comprises a gasket 200. In the illustratedembodiment, gasket 200 comprises a plurality of holes 201-209. Each holeis configured to receive a corresponding barrel located on bulk fluidsplate 100. Accordingly, first hole 201 is configured to receive firstbarrel 111; second hole 202 is configured to receive second barrel 112;third hole 203 is configured to receive third barrel 113; fourth hole204 is configured to receive fourth barrel 114; fifth hole 205 isconfigured to receive fifth barrel 115; sixth hole 206 is configured toreceive sixth barrel 116; seventh hole 207 is configured to receiveseventh barrel 117; eighth hole 208 is configured to receive eighthbarrel 118; and ninth hole 209 is configured to receive sampledispensing barrel 119. In specific embodiments, gasket 200 may beovermolded to bulk fluids plate 100.

In other embodiments not shown, a plurality of gaskets may be provided,such as equal in number to and corresponding to each barrel of bulkfluids plate 100.

FIGS. 11-19 illustrate an additional embodiment of a system 445 that canbe used for fluid assay analysis. In this embodiment, system 445comprises a fluid storage container 410, as well as other componentsdescribed more fully below. In this embodiment fluid storage container410 is comprised of a bulk fluids plate 427 with a syringe barrel array490, a plurality of pistons 431-436, sample input caps 438, 439,pierceable seals 419, and a gasket 485. Instead of blisters, thisembodiment employs pistons 431-436 to provide the positive displacementused to transfer the reagents and other fluids from fluid storagecontainer 410 to an analysis cartridge 420, described more fully below.This embodiment also utilizes coupling members 475, 465 to apply themechanical force required to maintain a substantial seal between fluidstorage container 410 and analysis cartridge 420 during fluid dispense.

Analysis Cartridge

As shown in FIGS. 1-3 and 8-10B, analysis cartridge 20 comprises aretention plate 500 configured to be coupled to bulk fluids plate 100.In the embodiments shown, retention plate 500 is configured to retain anassay surface, such as PCB 800, on the bottom side of retention plate500. In some embodiments, PCB 800 may comprise an electrowettingsurface, one or more heating elements, one or more microchannels, orsome combination of these features.

In the illustrated embodiment of FIGS. 1-10B, retention plate 500comprises a plurality of slots 510 that are configured to receivefastening tabs 182 and engagement tabs 184 of bulk fluids plate 100.

The embodiment shown in FIGS. 11-19 utilizes a different configurationfor retention plate 470. In this embodiment, retention plate 470comprises a pair of threaded holes 471, 472 into which coupling members475, 476 are inserted.

Referring back to the embodiment of FIGS. 1-10B, retention plate 500further comprises bosses protruding from its top side that correspond toand are configured to sealably mate with the barrels of container 10. Inthe illustrated embodiment, the bosses are configured to receive thebarrels of container 10. The bosses may be characterized as protrusionsin various embodiments. Each boss (discussed with reference to FIG. 8below) comprises a floor configured to limit the travel of itscorresponding barrel of container 10 and a piercing element, such as alance, disposed on and extending from the floor. In the illustratedembodiment, the lances are depicted as pointed or sharp. However, inother embodiments the piercing elements may be blunted, rounded, orflat-topped. Each piercing element is configured to pierce acorresponding seal located on a barrel of container 10.

Furthermore, in the illustrated embodiment, the bosses are depicted asbeing substantially cylindrical in shape; in other words, the bosseshave a substantially circular cross section. In other embodiments thebosses may not be cylindrical and may have substantially triangular,square, pentagonal, hexagonal, heptagonal, octagonal, or other polygonalcross sections, may be elliptical or partially rounded, or may haveirregular cross sections. Each boss and corresponding barrel isconfigured to sealably mate with each other such that liquid can movefrom the barrel to the boss with substantially no liquid leaking out. Inthe depicted embodiment, this configuration is achieved by the bosshaving a chamber configured to receive a barrel. In other embodiments,this could also be achieved by the barrel having a channel that isconfigured to receive a boss. In at least some such other embodiments,the top edge of the boss could act as a piercing element (and thus wouldbe one example of a piercing element associated with the boss), and thebottom of the boss's chamber could comprise a conduit through whichliquid may pass that enters the chamber after flowing from the piercedbarrel; in at least some other such embodiments, the boss could includea piercing element that extends upwardly from the bottom of the bosschamber to a location above the top edge of the boss (such a piercingelement being yet another example of a piercing element associated withthe boss), such that the piercing element is the first structure of theboss to contact the pierceable seal of the barrel. In embodiments wherethe barrel is configured to receive the boss, the shapes of the barrelchannel and the outside of the boss could each be tapered and configuredto fit tightly against each other to effect a substantial seal (wheresuch taper decreases in size as the relevant protrusion extends from therelevant plate).

In the illustrated embodiment, there are nine bosses that correspond tonine barrels—the eight reagent barrels 101-108 and one sample inputbarrel 109 of container 10. As shown in FIG. 8, first boss 501 comprisesa first chamber 541 having a first floor 521 and a first lance 531,which extends upward from the first floor and is a feature shared byeach of the floors and lances discussed herein. Second boss 502comprises a second chamber 542 having a second floor 522 and a secondlance 532. Third boss 503 comprises a third chamber 543 having a thirdfloor 523 and a third lance 533. Fourth boss 504 comprises a fourthchamber 544 comprising a fourth floor 524 and a fourth lance 534. Fifthboss 505 comprises a fifth chamber 545 having a fifth floor 525 and afifth lance 535. Sixth boss 506 comprises a sixth chamber 546 having asixth floor 526 and a sixth lance 536. Seventh boss 507 comprises aseventh chamber 547 having a seventh floor 527 and a seventh lance 537.Eighth boss 508 comprises an eighth chamber having an eighth floor 528and an eighth lance 538. Ninth boss 509 comprises a ninth chamber 549having a ninth floor 529 and a ninth lance 539.

A cross section detail view of an embodiment of first boss 501 is shownin FIGS. 10A and 10B. In the embodiments shown, first lance 531 is aconfigured to pierce seal 301 of first barrel 101. In preferredembodiments first lance 531 is about 0.150″ tall (that is, the distancefrom floor 521 to tip of lance 531 is about 0.150″). In otherembodiments, first lance 531 may be between about 0.150″ and about0.250″ tall. First lance 531 comprises a first conduit 561 in fluidcommunication with first dispensing reservoir 551. First conduit 561 isan example of a channel in (fluid) communication with first boss 501.First dispensing reservoir 551 is configured to deliver a volume offluid to PCB 800. Other bosses 502-509, lances 532-539, and dispensingreservoirs 552-559 comprise similar features that function in a similarmanner, which can be understood by a person of ordinary skill who refersto these figures. First lance 531 is an example of a piercing elementassociated with boss 501.

The illustrated embodiment of analysis cartridge 20 further comprises aplurality of oil-loading bosses 512 that are configured to be in fluidcommunication with PCB 800 and through which oil may be delivered to PCB800. The illustrated embodiment of analysis cartridge 20 also compriseliophylized reagent bosses 511 that are configured to be in fluidcommunication with PCB 800 and through which liophylized reagent may bedelivered to PCB 800.

In the illustrated embodiment, retention plate 500 further comprises aplurality of vents 540 in fluid communication with PCB 800 andconfigured to vent gas, such as air from PCB 800 during use. Vents 540are covered with membrane 300 which is configured to allow air to passbut not liquid. In the illustrated embodiment, membrane 300 is anadhesive-backed Versapor 800 hydrophobic nylon mesh.

Embodiments of the Present Systems and Methods

Embodiments of the present methods of using the illustrated embodimentof container 10 and cartridge 20 will be discussed with reference toFIGS. 1, 2, 10A and 10B.

In certain embodiments, sample input cap 151 may be removed fromcontainer 100 and a liquid sample introduced through sample inputchannel 152 into sample blister 153. Sample input cap 151 may then bereplaced. In certain embodiments, sample input cap 151 may be removedand replaced manually by a user; in other embodiments, sample input cap151 may be removed and replaced in an automated fashion.

Container 10 may be coupled to cartridge 20 by aligning fastening tabs182 and engagement tabs 184 of container 10 with slots 580 on cartridge20. In a fastening step, force is applied to container 10, cartridge 20,or both, such that fastening tabs 182 slide into the corresponding slots580 and container 10 is fastened to cartridge 20. Engagement tabs 184remain outside their corresponding slots 580 and are not directlyengaged with cartridge 20. None of the pierceable seals are pierced.This is known as the first stage, the first state, the fastened stage,or the fastened state.

As shown in FIG. 10A, at the fastened stage, the lances have not piercedthe seals of the corresponding barrels. However, barrel 111 has begun toengage boss 501.

In an engaging step, further force is applied to container 10, cartridge20, or both, such that engagement tabs 184 slide into the correspondingslots 580. This is known as the second stage, the engaged stage, thesecond state, or the engaged state.

As shown in FIG. 10B, at the engaged stage, the lances have pierced theseals of the barrels. For example, first lance 531 has pierced the seal301 located on first barrel 111. At the engaged stage, first barrel 111has traveled such that it is adjacent to and contacting first floor 521of first boss 501. Gasket 200 has traveled such that it is adjacent toand sealably contacting a portion of boss 501. In the engaged state,system 5 is configured such that gasket 200 will prevent substantiallyany fluid from leaking out of container 10 or cartridge 20. Morespecifically, such a configuration is achieved at least in part by theconfiguration of first barrel 111 contacting first floor 521, firstlance 531 being inside first reagent channel 121 such that first conduit561 is in fluid communication with first reagent reservoir 101, andfirst gasket 200 sealably engaging a portion of first barrel 111 andfirst boss 501 such that substantially any fluid is prevented fromleaking out of container 10 or cartridge 20 (that is, a substantiallyleak-proof seal is formed between container 10 and cartridge 20).

In certain embodiments, the fastening step may be accomplished manuallywhile the engaging step may be performed by an analysis deviceconfigured to receive container 10 and cartridge 20. In otherembodiments, both the fastening and engaging steps may be performed byan analysis device configured to receive container 10 and cartridge 20.In still other embodiments, both the fastening step and the engagingstep may be performed manually.

In a preferred embodiment, container 10 and cartridge 20 are introducedinto an instrument at the fastened stage. For example, the instrumentmay comprise a Luminex MAGPIX® multiplexing platform (available fromLuminex Corp., Austin, Tex.), though other suitable multiplexing orassay preparation instruments may be used.

Once introduced into the instrument, force is applied to container 10,cartridge 20, or both such that container 10 and cartridge 20 are in theengaged stage such that each lance 531-539 pierces the foil 301-309 onthe corresponding barrel 111-119.

Frangible lysis blister 161 is actuated, releasing the lysis buffercontained within. The lysis buffer rehydrates the sample control locatedin sample control reservoir 160. In addition, the lysis buffer travelsinto sample reservoir 150, lysing the sample.

In some embodiments, fourth flexible blister 134 and fifth flexibleblister 135 are actuated, which dispense the oil contained within fourthreagent reservoir 134 and fifth reagent reservoir 135 to cartridge 20,thereby displacing any air contained within cartridge 20.

Then, in some embodiments, first flexible blister 131, second flexibleblister 132, third flexible blister 133, sixth flexible blister 136,seventh flexible blister 137, and eighth flexible blister 138 areactuated to dispense the fluid contained within each of thecorresponding reagent reservoirs 101, 102, 103, 106, 107, and 108 todispensing reservoirs 531, 532, 533, 536, 537, and 538 in cartridge 20.

In some embodiments, sample blister 153 is then engaged, distributingthe lysed sample through sample dispensing channel 192 to cartridge 20.

These steps may be performed in the order listed in some embodiments,but may not be in other embodiments. Moreover, in other embodiments, notall the steps discussed above are performed. For example, fewer than allflexible blisters may be actuated.

An alternative embodiment comprising system 445 including fluid storagecontainer 410 and analysis cartridge 420 will be discussed further withreference to FIGS. 11-19.

In the embodiment shown, sample input caps 438, 439 may be removed frompistons 431, 434 and a liquid sample introduced into the samplereservoirs 451 and 454. In certain embodiments, sample input caps 438and 439 may be removed and replaced manually by a user; in otherembodiments, sample input caps 438, 439 may be removed and replaced inan automated fashion. As shown in FIG. 18, piston 431 may comprise aseal 437 extending around the circumference of piston 431. In certainembodiments, seal 437 may be an elastomer seal, and in specificembodiments, seal 437 may be configured as an O-ring.

Container 410 may be coupled to cartridge 420 by aligning couplingmembers 475, 476 on container 410 to threaded holes 471, 472 oncartridge 420. In a fastening step, torque can be applied to couplingmembers 475, 476 to thread coupling members 475, 476 into threaded holes471, 472. In this scenario seals 419 (also visible in FIG. 17) arepierced as assembled in FIG. 14. Additionally gasket 485 has traveledsuch that it is adjacent to and sealably contacting a raised feature 486on the floor of the cartridge 420.

In this state, system 445 is configured such that gasket 485 willprevent substantially any fluid from leaking out of container 410 orcartridge 420. More specifically, such a configuration is achieved atleast in part by the configuration of first protrusion 411 contactingfirst floor 403, first lance 491 being inside first reagent channel 421such that first conduit 492 is in fluid communication with samplereservoir 455, and 485 sealably engaging a portion of first protrusion411 and first boss 401 such that substantially any fluid is preventedfrom leaking out of container 410 or cartridge 420 (that is, asubstantially leak-proof seal is formed between container 410 andcartridge 420). As shown in FIG. 19, gasket 485 may comprise a pluralityof apertures or holes 484 corresponding with protrusions 411-416.Apertures 484 allow first reagent channel 421 to be in fluidcommunication with first conduit 492 after first protrusion 411 piercesseal 419.

Referring now to FIGS. 16 and 17, bulk fluids plate 427 is shown in aninverted perspective view. In this view, protrusions, 411-416 arevisible, as well as a portion of syringe barrel array 490. In FIG, 19,pierceable seals 419 are shown coupled to protrusions 411-416. Inaddition, coupling members 474, 476 are also shown coupled to bulkfluids plate 427 in FIG. 19. For purposes of clarity, it is understoodthat not all elements are labeled in all of the figures.

In one embodiment, container 410 and cartridge 420 are introduced intoan instrument at the fastened stage. For example, the instrument maycomprise a Luminex MAGPIX® multiplexing platform (available from LuminexCorp., Austin, Tex.), though other suitable multiplexing or assaypreparation instruments may be used.

After container 410 and cartridge 420 are loaded into the appropriateinstrument, oil piston 436 can be actuated to release oil from reservoir461 into cartridge 420 thereby displacing air within cartridge 420.

In some embodiments, sample pistons 431 and 434 are actuated, which candispense samples from sample reservoirs 455 and 458 into cartridge 420.In addition, pistons 432 and 433 can be actuated to dispense reagentscontained within each reagent reservoirs 456 and 457 into cartridge 20.Furthermore, piston 435 can be actuated to dispense magnetic particlesfrom reservoir 459 into cartridge 420.

These steps may be performed in the order listed in some embodiments,but may not be in other embodiments. Moreover, in other embodiments, notall the steps discussed above are performed. For example, fewer than allpistons may be actuated. It is understood that the embodiments of FIGS.11-19 may comprise additional features equivalent to those shown anddescribed in the discussion of the embodiments of FIGS. 1-10B.

Materials of Disclosed Embodiments

Non-limiting materials used to construct the illustrated embodiments ofthe present systems, containers, cartridges, and elements of these arediscussed below. Other suitable materials known to a person of ordinaryskill in the art may be used instead.

In the illustrated embodiment, bulk fluids plate 100 comprisespolycarbonate. In other embodiments bulk fluids plate 100 may compriseother semi-rigid plastics or hard plastics, which may includepolyurethanes, polyesters, epoxy resins and phenolic resins;polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), andpolyethylene terephthalate (PET or PETE). In still other embodiments,bulk fluids plate 100 may comprise one or more metals, such as aluminum.

In the illustrated embodiment, retention plate 500 comprisespolycarbonate. In other embodiments retention plate 500 may compriseother semi-rigid plastics or hard plastics, which may includepolyurethanes, polyesters, epoxy resins and phenolic resins;polyethylene (PE), polypropylene (PP) and polyvinyl chloride (PVC),polyethylene terephthalate (PET or PETE). In still other embodiments,retention plate 500 plate may comprise metals such as aluminum.

Frangible lysis blister 161 is a Thinxxs 500 uL frangible blister in theembodiment shown. However, lysis blister may comprise other flexibleand/or frangible polymers in other embodiments.

Blister layer 130, flexible blisters 131-139, and sample blister 153comprise LDPE in the illustrated embodiment. In other embodiments, theseblisters may comprise other flexible polymers.

Adhesive layers 140 and 141 comprise adhesive transfer tape (3M, 9485PCadhesive transfer tape) in the illustrated embodiment, though otherforms of adhesive may be used.

In the illustrated embodiment, gasket 200 comprises polyurethane. Inspecific embodiments, gasket 200 comprises a 0.06″ sheet of polyurethanethat has been die-cut and coupled to bulk fluids plate 100 with adhesivetransfer tape. Other suitable materials for gasket 200, such aspolymers, may be used.

Seals 301-309 in the illustrated embodiments comprise aluminum foilbacked with bi-axially oriented polypropylene film (BOPP) (such as0.001″ aluminum foil backed with 0.002″ BOPP film).

It should be understood that the present devices and methods are notintended to be limited to the particular forms disclosed. Rather, theyare to cover all modifications, equivalents, and alternatives fallingwithin the scope of the claims. For example, certain embodiments of thecontainer 10 and cartridge 20 discussed above are shown configured foruse with an assay preparation module. However, container 10 andcartridge 20 are suitable for use in any small space where precisedispensation of fluids in a specified order may be required.

The above specification and examples provide a complete description ofthe structure and use of an exemplary embodiment. Although certainembodiments have been described above with a certain degree ofparticularity, or with reference to one or more individual embodiments,those skilled in the art could make numerous alterations to thedisclosed embodiments without departing from the scope of thisinvention. As such, the illustrative embodiment of the present devicesis not intended to be limited to the particular forms disclosed. Rather,they include all modifications and alternatives falling within the scopeof the claims, and embodiments other than the one shown may include someor all of the features of the depicted embodiment. For example,components may be combined as a unitary structure and/or connections maybe substituted. As another example, one of ordinary skill in the artwould understand that, in alternate embodiments, fastening tabs 182 andengagement tabs 184 may be located on cartridge 20 while slots 580 maybe located on container 10. Further, where appropriate, aspects of anyof the examples described above may be combined with aspects of any ofthe other examples described to form further examples having comparableor different properties and addressing the same or different problems.Similarly, it will be understood that the benefits and advantagesdescribed above may relate to one embodiment or may relate to severalembodiments.

The claims are not to be interpreted as including means-plus- orstep-plus-function limitations, unless such a limitation is explicitlyrecited in a given claim using the phrase(s) “means for” or “step for,”respectively.

We claim:
 1. A method comprising: (a) adding a volume of a sample to asample reservoir of a system, wherein the system comprises: (1) a firstplate comprising: (i) a first side; (ii) a second side; (iii) a samplereservoir; (iv) a first port; (v) a channel connecting the samplereservoir and the first port; (vi) a sample dispensing channel openingonto the second side of the first plate, wherein the sample dispensingchannel is in fluid communication with the sample reservoir; (2) afrangible blister coupled to the first side of the plate and coveringthe first port; (3) a second plate comprising: (i) a first side; (ii) asecond side; (iii) a conduit passing through the second plate from thefirst side of the second plate to the second side of the second plate;wherein the first side of the second plate is coupled to the second sideof the first plate; and (4) a printed circuit board retained on thesecond side of the second plate; wherein the printed circuit boardcomprises an electrowetting surface; (b) lysing the sample in the firstplate, wherein the lysing comprises actuating the frangible blister totransfer a lysis buffer from the frangible blister to the samplereservoir via the first port and the channel connecting the samplereservoir and the first port; and (c) transferring the lysed sample fromthe first plate to the electrowetting surface of the printed circuitboard via the conduit passing through the second plate.
 2. The method ofclaim 1, wherein the actuating of the frangible blister is performedmanually.
 3. The method of claim 1, wherein the printed circuit boardcomprises one or more heating elements.
 4. The method of claim 1, wherethe first side of the second plate is coupled to the second side of thefirst plate prior to adding the volume of sample to the sample reservoir5. The method of claim 1, wherein: the first plate comprises: aplurality of protrusions extending from the second side of the firstplate; and a plurality of pierceable seals, where each pierceable sealin the plurality of pierceable seals is coupled to one of theprotrusions in the plurality of protrusions extending from the secondside of the first plate; and the first side of the second plate iscoupled to the second side of the first plate in a first stage with thepierceable seals intact prior to adding the volume of sample to thesample reservoir unit.
 6. The method of claim 5, further comprisingcoupling the first side of the second plate to the second side of thefirst plate in a second stage, wherein at least one of the pierceableseals pierced.
 7. The method of claim 1, wherein the first side of thesecond plate is coupled to the second side of the first plate afteradding the volume of sample to the sample reservoir.
 8. The method ofclaim 1, wherein: the second side of the first plate comprises aplurality of reagent reservoir units, each of which comprises a fluiddisplacement mechanism and a fluid volume; and the first side of thesecond plate comprises a plurality of bosses; and further comprisingactuating the fluid displacement mechanism of at least one of theplurality of reagent reservoir units to deliver the volume of fluid inthe reagent reservoir unit to the electrowetting surface of the printedcircuit board via a conduit of one of the bosses.
 9. The method of claim8, further comprising actuating two or more of the fluid displacementmechanisms simultaneously.
 10. The method of claim 8, further comprisingactuating two or more of the fluid displacement mechanisms sequentially.11. The method of claim 8, wherein at least one of the plurality ofreagent reservoir units comprises a volume of oil, and the fluiddisplacement mechanism of the at least one reagent reservoir unitcomprising a volume of oil is actuated before actuating the fluiddisplacement mechanism of any of the other reagent reservoir units andbefore transferring the lysed sample from the first plate to theelectrowetting surface of the printed circuit board.